Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4249

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Linkage Analysis Using the NaOH Methylation Method

David Oxley, Graeme Currie, and Antony Bacic

This protocol was adapted from "Linkage Analysis Using the NaOH Methylation Method," contributed by David Oxley, Graeme Currie, and Antony Bacic, Chapter 25, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2004.

The first 100 words of the full text of this article appear below.


INTRODUCTION

Linkage analysis provides information on sugar type, ring size, and substitution positions for each monosaccharide. The method in this protocol, using NaOH as the base, is one of the simpler linkage analysis methods. It requires approximately 1-5 µg of carbohydrate.


MATERIALS

Reagents

caution Acetic acid, glacial

caution Acetic acid (5%) in methanol

caution Acetic anhydride, glacial

caution Dichloromethane (DCM)

caution Dimethylsulfoxide (DMSO)

Glycoprotein, proteoglycan, or glycan sample of interest, vacuum-dried (1-5 µg)

Methanol, anhydrous

caution Methyl iodide (100%)

myo-Inositol (20 mg/ml in H2O)

myo-Inositol is used as an internal standard. If myo-inositol is suspected to be in the sample (e.g., GPI anchors), substitute scyllo. . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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